Inhaled nitric oxide reverses PAF-dependent bronchoconstriction in the pig

In six anesthetized, paralyzed, mechanically ventilated pigs we evaluated the respiratory effects of inhaled nitric oxide (NO) (80 ppm in O₂) under control conditions and after platelet-activating factor (PAF) administration (50 ng/kg, i.v.). PAF was also administered to the same pigs after pretreatment with indomethacin (3 mg/kg, i.v.). The mechanical properties of the respiratory system were evaluated by the rapid airway occlusion technique. With this technique the overall respiratory resistances, the airway resistances, and the additional resistances of respiratory system and lung can be evaluated. The results show that NO inhaled by the pig at 80 ppm for 6 min under control conditions reduced static and dynamic elastances of the respiratory system and lung and pulmonary arterial pressure, without modifying bronchomotor tone. Therefore, NO reduced the PAF-dependent changes in resistances and in static and dynamic elastances of the respiratory system and lung. The modest change in elastances caused by PAF in pigs pretreated with indomethacin was reduced by NO inhalation, which also has a mild bronchodilatory effect. The changes in elastances appear to be correlated with the pulmonary vasodilator activity of inhaled NO.     

Inhaled Nitric Oxide Reverses Vascular and Respiratory Effects of ET-1 and PAF in Pigs

In anaesthetized paralysed, mechanically ventilated pigs, the vascular and respiratory effects of 80 ppm nitric oxide (NO) inhaled for 6 min were evaluated. To evoke different levels of smooth muscle contraction ET-1 or PAF, mediators involved in pulmonary disorders, were used. In control conditions, inhaled NO caused selective pulmonary vasodilatation without affecting respiratory resistances. This pulmonary vascular activity influenced the distensibility of the respiratory system and decreased inspiratory work. ET-1 administration significantly increased pulmonary arterial pressure and modestly changed mechanical properties of the respiratory system, while PAF caused potent vasoconstriction and bronchoconstriction associated with a marked change in volume-pressure relationship. In both cases, the changes in vascular and mechanical properties of the respiratory system increased inspiratory work. The vascular and respiratory activities of inhaled NO were correlated with preconstriction levels. The data show that the combination of vascular and respiratory effects improves pulmonary function, suggesting that inhalation of NO is a possible therapeutic approach for obstructive and inflammatory pulmonary diseases.     

Inhaled nitric oxide induced NOS inhibition and rebound pulmonary hypertension: a role for superoxide and peroxynitrite in the intact lamb

Previous in vivo studies indicate that inhaled nitric oxide (NO) decreases nitric oxide synthase (NOS) activity and that this decrease is associated with significant increases in pulmonary vascular resistance (PVR) upon the acute withdrawal of inhaled NO (rebound pulmonary hypertension). In vitro studies suggest that superoxide and peroxynitrite production during inhaled NO therapy may mediate these effects, but in vivo data are lacking. The objective of this study was to determine the role of superoxide in the decrease in NOS activity and rebound pulmonary hypertension associated with inhaled NO therapy in vivo. In control lambs, 24 h of inhaled NO (40 ppm) decreased NOS activity by 40% (P<0.05) and increased endothelin-1 levels by 64% (P<0.05). Withdrawal of NO resulted in an acute increase in PVR (60.7%, P<0.05). Associated with these changes, superoxide and peroxynitrite levels increased more than twofold (P<0.05) following 24 h of inhaled NO therapy. However, in lambs treated with polyethylene glycol-conjugated superoxide dismutase (PEG-SOD) during inhaled NO therapy, there was no change in NOS activity, no increase in superoxide or peroxynitrite levels, and no increase in PVR upon the withdrawal of inhaled NO. In addition, endothelial NOS nitration was 18-fold higher (P<0.05) in control lambs than in PEG-SOD-treated lambs following 24 h of inhaled NO. These data suggest that superoxide and peroxynitrite participate in the decrease in NOS activity and rebound pulmonary hypertension associated with inhaled NO therapy. Reactive oxygen species scavenging may be a useful therapeutic strategy to ameliorate alterations in endogenous NO signaling during inhaled NO therapy.     

Nitric oxide modulation of the crystallogenic properties of a biological fluid

A comparative analysis of the influence of different nitric oxide forms on the character of the dehydration structuring of human serum samples was carried out. The effects of an NO-containing gas flow that was generated by a Plazon device (800 and 80 ppm), an experimental NO generator (20, 50, 75, and 100 ppm), as well as by glutathione-containing dinitrosyl iron complexes (3 mM/L) were investigated using 15 healthy donors. The influence of endogenous sodium on serum crystallization of intact and NO treated blood samples was evaluated. It was found that the effect of NO on the crystallogenic properties of blood serum is directly determined by its concentration and form (free or bound), as well as by the presence of reactive oxygen species in a gas flow. The most pronounced stimulatory effect was observed for the bound NO form, dinitrosyl iron complexes with glutathione ligands. Low NO concentrations modulated the crystallogenic properties of blood serum, while the best stimulatory effect was demonstrated by the gas flow that contained 20 ppm nitric oxide. In contrast, high NO concentrations (800 ppm) inhibited the crystallogenic activity of the biological medium due to an increase in the destruction of structural elements by many times, which lead to the formation of an additional band in the marginal zone of the microscope specimen.     

Endothelial alterations during inhaled NO in lambs with pulmonary hypertension: implications for rebound hypertension

Clinically significant increases in pulmonary vascular resistance (PVR) have been noted upon acute withdrawal of inhaled nitric oxide (iNO). Previous studies in the normal pulmonary circulation demonstrate that iNO increases endothelin-1 (ET-1) levels and decreases endogenous nitric oxide synthase (NOS) activity, implicating an endothelial etiology for the increase in resistance upon iNO withdrawal. However, the effect of iNO on endogenous endothelial function in the clinically relevant pulmonary hypertensive circulation is unknown. The objective of this study was to determine the effects of iNO on endogenous NO-cGMP and ET-1 signaling in lambs with preexisting pulmonary hypertension secondary to increased pulmonary blood flow. Eight fetal lambs underwent in utero placement of an aortopulmonary vascular graft (shunt lambs). After delivery (4 wk), the shunt lambs were mechanically ventilated with iNO (40 ppm) for 24 h. After 24 h of inhaled NO, plasma ET-1 levels increased by 34.8% independently of changes in protein levels (P < 0.05). Contrary to findings in normal lambs, total NOS activity did not decrease during iNO. In fact, Western blot analysis demonstrated that tissue endothelial NOS protein levels decreased by 43% such that NOS activity relative to protein levels actually increased during iNO (P < 0.05). In addition, the beta-subunit of soluble guanylate cyclase decreased by 70%, whereas phosphodiesterase 5 levels were unchanged (P < 0.05). Withdrawal of iNO was associated with an acute increase in PVR, which exceeded baseline PVR by 45%, and a decrease in cGMP concentrations to levels that were below baseline. These data suggest that the endothelial response to iNO and the potential mechanisms of rebound pulmonary hypertension are dependent upon the underlying pulmonary vasculature.     

Neurokinins and inflammatory cell iNOS expression in guinea pigs with chronic allergic airway inflammation

In the present study we evaluated the role of neurokinins in the modulation of inducible nitric oxide synthase (iNOS) inflammatory cell expression in guinea pigs with chronic allergic airway inflammation. In addition, we studied the acute effects of nitric oxide inhibition on this response. Animals were anesthetized and pretreated with capsaicin (50 mg/kg sc) or vehicle 10 days before receiving aerosolized ovalbumin or normal saline twice weekly for 4 wk. Animals were then anesthetized, mechanically ventilated, given normal saline or N(G)-nitro-l-arginine methyl ester (l-NAME, 50 mg/kg ic), and challenged with ovalbumin. Prechallenge exhaled NO increased in ovalbumin-exposed guinea pigs (P < 0.05 compared with controls), and capsaicin reduced this response (P < 0.001). Compared with animals inhaled with normal saline, ovalbumin-exposed animals presented increases in respiratory system resistance and elastance and numbers of total mononuclear cells and eosinophils, including those expressing iNOS (P < 0.001). Capsaicin reduced all these responses (P < 0.05) except for iNOS expression in eosinophils. Treatment with l-NAME increased postantigen challenge elastance and restored both resistance and elastance previously attenuated by capsaicin treatment. Isolated l-NAME administration also reduced total eosinophils and mononuclear cells, as well as those cells expressing iNOS (P < 0.05 compared with ovalbumin alone). Because l-NAME treatment restored lung mechanical alterations previously attenuated by capsaicin, NO and neurokinins may interact in controlling airway tone. In this experimental model, NO and neurokinins modulate eosinophil and lymphocyte infiltration in the airways.     

The features of lipid peroxidation and the antioxidant system of blood under the influence of different concentrations of nitric oxide in a chronic experiment

This paper presents the results of the study of oxidative metabolism of the blood of intact animals subjected to prolonged exposure to nitric oxide at concentrations of 20, 50, and 100 ppm. The experiment was carried out with Wistar rats. NO inhalation was performed for 30 days. The state of blood oxidative metabolism was evaluated after inhalation and a 30-day-long recovery period after discontinuation of NO oxidative stress. The intensity of lipid peroxidation was studied in plasma and erythrocytes by induced biochemiluminescence and the measurement of the level of malondialdehyde. The activity of superoxide dismutase was determined in the hemolysate of erythrocytes. It was established that the optimal dose of inhaled NO is 20 ppm: a maximum increase in the total antioxidant activity after 30 days and normalization of lipid peroxidation in the blood after the completion of the recovery period were observed at this concentration. High concentrations of nitric oxide (50 and 100 ppm) initiated lipid peroxidation in erythrocytes and plasma after discontinuation of NO oxidative stress (after the completion of the recovery period) thus enhancing catalytic properties of superoxide dismutase.     

Inhaled nitric oxide decreases infarction size and improves left ventricular function in a murine model of myocardial ischemia-reperfusion injury

To learn whether nitric oxide (NO) inhalation can decrease myocardial ischemia-reperfusion (I/R) injury, we studied a murine model of myocardial infarction (MI). Anesthetized mice underwent left anterior descending coronary artery ligation for 30, 60, or 120 min followed by reperfusion. Mice breathed NO beginning 20 min before reperfusion and continuing thereafter for 24 h. MI size and area at risk were measured, and left ventricular (LV) function was evaluated using echocardiography and invasive hemodynamic measurements. Inhalation of 40 or 80 ppm, but not 20 ppm, NO decreased the ratio of MI size to area at risk. NO inhalation improved LV systolic function, as assessed by echocardiography 24 h after reperfusion, and systolic and diastolic function, as evaluated by hemodynamic measurements 72 h after reperfusion. Myocardial neutrophil infiltration was reduced in mice breathing NO, and neutrophil depletion prevented inhaled NO from reducing myocardial I/R injury. NO inhalation increased arterial nitrite levels but did not change myocardial cGMP levels. Breathing 40 or 80 ppm NO markedly and significantly decreased MI size and improved LV function after ischemia and reperfusion in mice. NO inhalation may represent a novel method to salvage myocardium at risk of I/R injury.     

Co-localisation of nitric oxide synthase and endothelin in the rat supraoptic nucleus

The co-localisation of neuronal nitric oxide synthase and endothelin-1 was studied in the rat supraoptic nucleus at the electron microscopy level. Double pre-embedding immunocytochemistry was performed using ExtrAvidin-horseradish peroxidase and immunogold–silver techniques. Immunoreactivities to neuronal nitric oxide synthase and endothelin-1 were co-localised in sub-populations of endocrine neurones (cell bodies) and dendrites. Double-labelled axon terminals making asymmetrical synapses on unlabelled dendrites were also observed. The findings are discussed in terms of the possible role and significance of nitric oxide and endothlin-1 in the hypothalamo-neurohypophysial system.     

Effects of acute and chronic nitric oxide inhibition in an experimental model of chronic pulmonary allergic inflammation in guinea pigs

Endogenously produced nitric oxide is a recognized regulator of physiological lung events, such as a neurotransmitter and a proinflammatory mediator. We tested the differences between chronic and acute nitric oxide inhibition by N(omega)-nitro-L-arginine methyl ester (L-NAME) treatment in lung mechanics, inflammation, and airway remodeling in an experimental asthma model in guinea pigs. Both acute and chronic L-NAME treatment reduced exhaled nitric oxide in sensitized animals (P < 0.001). Chronic L-NAME treatment increased baseline and maximal responses after antigen challenge of respiratory system resistance and reduced peribronchial edema and mononuclear cells airway infiltration (P < 0.05). Acute administration of L-NAME increased maximal values of respiratory system elastance and reduced mononuclear cells and eosinophils in airway wall (P < 0.05). Chronic ovalbumin exposure resulted in airway wall thickening due to an increase in collagen content (P < 0.005). Chronic nitric oxide inhibition increased collagen deposition in airway wall in sensitized animals (P < 0.05). These data support the hypothesis that in this model nitric oxide acts as a bronchodilator, mainly in proximal airways. Furthermore, chronic nitric oxide inhibition was effective in reducing edema and mononuclear cells in airway wall. However, airway eosinophilic inflammation was unaltered by chronic L-NAME treatment. In addition, nitric oxide inhibition upregulates collagen deposition in airway walls.     

Could nasal nitric oxide help to mitigate the severity of COVID-19?

The nasal cavity and turbinates play important physiological functions by filtering, warming and humidifying inhaled air. Paranasal sinuses continually produce nitric oxide (NO), a reactive oxygen species that diffuses to the bronchi and lungs to produce bronchodilatory and vasodilatory effects. Studies indicate that NO may also help to reduce respiratory tract infection by inactivating viruses and inhibiting their replication in epithelial cells. In view of the pandemic caused by the novel coronavirus (SARS-CoV-2), clinical trials have been designed to examine the effects of inhaled nitric oxide in COVID-19 subjects. We discuss here additional lifestyle factors such as mouth breathing which may affect the antiviral response against SARS-CoV-2 by bypassing the filtering effect of the nose and by decreasing NO levels in the airways. Simple devices that promote nasal breathing during sleep may help prevent the common cold, suggesting potential benefits against coronavirus infection. In the absence of effective treatments against COVID-19, the alternative strategies proposed here should be considered and studied in more detail.     

一种简易的小动物一氧化氮吸入系统的研制

作者自行设计一套小动物一氧化氮吸入系统,该系统主要包括有机玻璃舱,采气泵,流量计,ＮＯ／Ｎ２混和气体,纯氧,氮氧化物分析仪,Ｏ２,ＣＯ２监测仪。实验过程中,将６－７只大鼠置于舱内分别吸入２０ｐｐｍ,４０ｐｐｍ,８０ｐｐｍ和ＮＯ８ｈ,每２０ｍｉｎ记录一次舱内ＮＯ,ＮＯ２,Ｏ２和ＣＯ２的浓度值。结果表明舱内的实测ＮＯ浓度与实验所设计的ＮＯ浓度相近,舱内氧浓度与空气中的氧浓度相似,ＮＯ和ＣＯ２最大浓度分     

Endothelin-1 response to mental stress in early ischemic lesions of the extremities due to systemic sclerosis

We studied circulating levels of endothelin-1, catecholamines and nitric oxide after a mental arithmetic test in 14 patients with early ischemic lesions of the extremities due to systemic sclerosis and slightly impaired peripheral vascular flow. The test induced an increase (P < 0.01) in blood pressure, heart rate, endothelin-1 and catecholamine levels, whereas it did not change the low basal levels of nitric oxide. In healthy subjects (n = 20) the test significantly (P < 0.01) decreased endothelin-1 without affecting nitric oxide. The low basal levels of nitric oxide and the high plasma concentration of endothelin-1 after psychological stress cannot be explained by an impaired release from the limited ischemic lesions alone. This suggests a diffuse microvascular derangement that aggravates the course of peripheral microvascular ischemic lesions.     

Erythropoietin acutely decreases airway resistance in the rat

While some experimental data suggest that erythropoietin (EPO) influences respiratory mechanics, reports on scientific trials are lacking. In the present work, respiratory mechanics were measured using the end-inflation occlusion method in control and EPO treated anaesthetised and positive-pressure ventilated rats. Causing an abrupt inspiratory flow arrest, the end-inflation occlusion method makes it possible to measure the ohmic airway resistance and the respiratory system elastance. It was found that EPO induces a significant decrement in the ohmic airway resistance, not noted in control animals, 20 and 30min after intraperitoneal EPO injection. The elastic characteristics of the respiratory system did not vary. Hypotheses about the mechanism (s) explaining these results were addressed. In particular, additional experiments have indicated that the decrement in airway resistance could be related to an increase in nitric oxide production induced by EPO. Spontaneous increments in plasmatic erythropoietin levels, such as those that take place in association with hypoxia and/or blood loss, appear to be related to the decrement in airway resistance, allowing pulmonary ventilation to increase without altering respiratory mechanics leading to deleterious increments in energy dissipation during breathing.     

Differential effects of inhaled nitric oxide and hyperoxia on pulmonary dysfunction in newborn guinea pigs

This study tested the hypothesis that inhaled nitric oxide (NO) and combined NO and hyperoxia will result in less pulmonary dysfunction and delay onset of respiratory signs compared with hyperoxia-exposed newborn guinea pigs (GPs). GPs were exposed to room air (n = 14), 95% O(2) (n = 36), 20 parts per million (ppm) NO (n = 14), or combined 20 ppm NO and 95% O(2) (NO/O(2), n = 13) for up to 5 days. Data evaluated included latency interval for onset of respiratory distress, pressure volume curves, lung histology, and bronchoalveolar lavage (BAL) polymorphonuclear cells (PMNs), proteolytic activity, and total protein. NO-exposed GPs did not develop respiratory distress and had no evidence of pulmonary dysfunction. O(2)-exposed GPs developed respiratory distress after 1-5 days (median 4.0) vs. 3-5 days (median 5.0) for NO/O(2) exposure (P < 0.05). BAL from O(2)-exposed GPs showed increased PMNs compared with NO/O(2)-exposed GPs. O(2)- and NO/O(2)-exposed GPs had comparable reduced lung volumes, lung histology, and increased BAL proteinase activity and total protein. In summary 1) O(2) exposure resulted in multiple measures of pulmonary dysfunction in newborn GPs, 2) 5-day exposure to NO produced no noticeable respiratory effects and pulmonary dysfunction, and 3) short-term exposure (相似文献   

Effects of inhaled nitric oxide at rest and during exercise in idiopathic pulmonary fibrosis

Patients with idiopathic pulmonary fibrosis (IPF) usually develop hypoxemia and pulmonary hypertension when exercising. To what extent endothelium-derived vasodilating agents modify these changes is unknown. The study was aimed to investigate in patients with IPF whether exercise induces changes in plasma levels of endothelium-derived signaling mediators, and to assess the acute effects of inhaled nitric oxide (NO) on pulmonary hemodynamics and gas exchange, at rest and during exercise. We evaluated seven patients with IPF (6 men/1 woman; 57 ± 11 yr; forced vital capacity, 60 ± 13% predicted; carbon monoxide diffusing capacity, 52 ± 10% predicted). Levels of endothelin, 6-keto-prostaglandin-F(1α), thromboxane B(2), and nitrates were measured at rest and during submaximal exercise. Pulmonary hemodynamics and gas exchange, including ventilation-perfusion relationships, were assessed breathing ambient air and 40 ppm NO, both at rest and during submaximal exercise. The concentration of thromboxane B(2) increased during exercise (P = 0.046), whereas levels of other mediators did not change. The change in 6-keto-prostaglandin-F(1α) correlated with that of mean pulmonary arterial pressure (r = 0.94; P < 0.005). Inhaled NO reduced mean pulmonary arterial pressure at rest (-4.6 ± 2.1 mmHg) and during exercise (-11.7 ± 7.1 mmHg) (P = 0.001 and P = 0.004, respectively), without altering arterial oxygenation or ventilation-perfusion distributions in any of the study conditions. Alveolar-to-capillary oxygen diffusion limitation, which accounted for the decrease of arterial Po(2) during exercise, was not modified by NO administration. We conclude that, in IPF, some endothelium-derived signaling molecules may modulate the development of pulmonary hypertension during exercise, and that the administration of inhaled NO reduces pulmonary vascular resistance without disturbing gas exchange.     

Secretory function of the endothelium as a factor of vascular tone regulation in the norm and in cardiovascular pathology

Endothelin-1 and nitric oxide are the most potent factors of the endothelium-derived substances. The factors play opposite roles in regulation of cardiovascular system, and their interaction underlies the balance of vasoconstrictor and vasodilator influences on vascular tone under normal conditions. In our experiments, changes in endothelin-1 blood concentration were associated with affected production of endogenous nitric oxide. The altered interrelationships between the endothelium-derived vasoactive substances may precede pathological shifts in the cardiovascular system.     

Generation of nitric oxide by human neutrophils

Human neutrophils were evaluated for their ability to generate nitric oxide. Neutrophils incubated with superoxide dismutase at 37 degrees C produce nitrite anion at a rate of 1.8 nmols/2 x 10(6) cells/30 min, providing indirect evidence of nitric oxide production. Incubation of the neutrophils with concentrations of serum-opsonized zymosan, N-formyl-methionyl-leucyl-phenylalanine, or phorbol myristate acetate sufficient to stimulate the respiratory burst and lysosomal enzyme release caused no additional nitrite anion production. Glass wool-adherent neutrophils exhibited a similar dissociation of nitrite anion production from the respiratory burst and lysosomal enzyme release. Direct evidence for nitric oxide production was also obtained using nitric oxide-specific chemiluminescence. These results demonstrate that human neutrophils are capable of generating nitric oxide.     

Improvement of respiratory function by bosentan during endotoxic shock in the pig.

We evaluated the role of endothelin-1 (ET-1) and the involvement of nitric oxide in cardiovascular and respiratory dysfunction, during endotoxic shock, in 18 anaesthetised, mechanically ventilated pigs, divided into three groups. Group 1 was i.v. infused with LPS (20 microg/Kg/h for 240 min). Group 2 was pre-treated with bosentan, a dual inhibitor of ET-1 receptors, and at 180 min of endotoxic shock, L-NAME (N(G)-nitro-L-arginine methyl ester, 10 mg/Kg), a non-selective inhibitor of NO synthases, was i.v. administered. Group 3 was infused with LPS and L-NAME was administered similarly to group 2. Results show that LPS caused systemic hypotension, pulmonary biphasic hypertension, decrease in compliance (C(rs)) and increase in resistance (R(max,rs)) of respiratory system. Bosentan completely abolished the pulmonary hypertension and the changes in C(rs)and R(max,rs). L-NAME does not affect the LPS-dependent changes in respiratory mechanics, but it worsens the cardiovascular effects, causing death of pigs. Pre-treatment with bosentan prevents this deleterious effect.Our study demonstrates that the LPS-dependent respiratory effects are mediated by ET-1, which, probably causing pulmonary oedema, is responsible for the decrease in C(rs)and the increase of R(max,rs).     

Transient injury to rat lung mitochondrial DNA after exposure to hyperoxia and inhaled nitric oxide

The effect of hyperoxia alone and in combination with inhaled nitric oxide (NO) on the integrity of lung mitochondrial DNA (mtDNA) in vivo was evaluated in Fischer 344 rats. PCR amplification of lung mtDNA using two sets of primers spanning 10.1 kb of the mtDNA revealed that inhalation of 20 ppm of NO in conjunction with hyperoxia (>95% O2) reduced the amplification of mtDNA templates by 10 +/- 1% and 26 +/- 3% after 24 h of exposure. The ability of mtDNA to amplify was not compromised in rats exposed to 80% O2, even in the presence of 20 ppm of inhaled NO. Surprisingly, exposure to >95% O2 alone for either 24 or 48 h did not compromise the integrity of mtDNA templates compared with air-exposed controls, despite evidence of genomic DNA injury. Interestingly, inhaling NO alone for 48 h increased mtDNA amplification by 12 +/- 2% to 21 +/- 7%. Injury to the lung mtDNA after exposure to >95% O2 plus 20 ppm of NO was transient as rats allowed to recover in room air after exposure displayed increased amplification, with levels exceeding controls by 20 +/- 3% to 29 +/- 4%. Increased amplification was not due to cellular proliferation or increased mitochondrial number. Moreover, the ratio of pulmonary mtDNA to genomic DNA remained the same between treatment groups. The results indicate that hyperoxia fails to induce significant injury to mtDNA, and whereas inhalation of NO with hyperoxia results in mtDNA damage, the lesions are rapidly repaired during recovery.